양송이배지로부터 분리한 인산가용화균 Burkholderia contaminans PSB-A와 Burkholderia ambifaria PSB-B의 단일접종 및 동시접종에 따른 인산가용화 능력의 시너지 효과 및 상추생육효과를 조사하였다. 인산캄슘이 함유된 NBRIP 액체배지에 분리균을 접종하고 해리된 가용인산 (soluble phosphorus)함량을 HPLC에 의해 분석하여 인산 가용화능을 측정한 결과, 배양 3일차에 동시접종구 166.3 μg mL -1 , B. contaminans 143.7 μg mL -1 , B. ambifaria 127.1 μg mL -1의 순으로 나타나 두 종의 인산가용화세균 간의 동시접종에 의한 시너지효과가 다소 관찰되었다. 배지 내 pH 와 잔류 glucose 함량 변화도 모든 접종구에서 배양 1 - 3일 후 pH 7.0에서 pH 4.0 수준으로 감소하였으며, glucose 함량은 초기 10 mg mL -1 에서 4.3 mg mL -1 수준으로 검출되어 인산가용화능과 거의 유사한 경향을 보였다. 배양여액의 유기산 분석결과 gluconic acid 약 12.3 mg mL -1 와 malic acid 약 7.9 mg mL -1 수준으로 가장 높게 검출되었으며, 결과적으로 두 세균이 분비하는 유기산이 배지의 pH를 감소시키고, 인산 가용화를 유도하는 주요 원인임을 확인할 수 있었다. 접종 4주 후 상추 생육차이를 조사한 결과, TCP를 첨가하지 않은 B. contaminans 접종구, B. ambifaria 접종구와 동시 접종구 에 비해 TCA 첨가한 모든 처리구에서 약 7-9% 수준의 생육증진효과가 보였으나, 단일접종과 동시접종구간에 유의수준의 차이를 확인할 수 없었다.

An auxin-producing bacterium Yangsong-1 was isolated from a button mushroom bed in Chung cheongnam-do. The strain Yangsong-1 was classified as a novel strain of Arthrobacter enclensis based on a chemotaxonomic and phylogenetic analysis. The isolated A. enclensis Yangsong-1 was confirmed to produce indole-3-acetic acid (IAA), which is one of the auxin hormones. When the concentration of IAA was assessed by HPLC quantity analysis, the maximum concentration of IAA, 152.903 mg L-1, was detected from the culture broth incubated in R2A medium containing 0.2% L-tryptophan for 48 h at 35oC. A negative relationship between IAA production and pH was estimated to show that the increase in IAA caused pH acidification of the culture. The effect of the supplement on L-tryptophan, a known precursor of IAA production, appeared to be at maximal production at 0.2% concentration and was rather reduced at concentration above 0.4%. To investigate the growth-promoting effects on the crops, the culture broth of A. enclensis Yangsong-1 was placed in water cultures and seed pots of mung beans and lettuce. In consequence, the adventitious root induction and root growth of mung beans and lettuce were 1.5 and 1.9 times higher, respectively, than those of the control.

충청남도 부여군 석성면 양송이 재배 농가에서 양송이 수확 후 배지로부터 토양을 채취하여 auxin(IAA) 생성능이 뛰어난 세균 Jopap-1 균주를 분리하였다. TLC 및 HPLC 분석을 통해 분리균이 생성한 IAA 농도를 확인한 결과, 0.1% L-tryptophan를 함유한 pH 7.0의 R2A broth 배지에 35℃, 48시간 배양 시 최대 생성농도는 96.05 mg L−1이었다. 생리적 특성 및 계통학적특성 분석을 통해 분리균은 Gram 음성 간균인 Klebsiella michiganensis Jopap-1로 동정되었다. 배양조건별 IAA의 생산능 비교 시, IAA 농도의 증가가 배양액의 pH 산성화에 기인함으로서 IAA 생성량과 pH 변화에는 부의 상관성이 있는 것으로 관측되었다. IAA 생성을 위한 전구물질로 알려진 L-tryptophan의 첨가효과는 0.1% 첨가 시 균의 생육 및 IAA 생성량이 최대이었으며, 0.2% 이상 고농도 첨가 배지에서는 오히려 IAA의 생성이 저해되었다. 또한 분리균에 의한 식물 생육 촉진 효과를 조사하기 위하여 수경재배 및 pot 재배를 통한 녹두발근 생검법과 상추발근 생검법을 수행한 결과, K. michiganensis Jopap-1의 배양액 접종 시 녹두발근 생검법에서는 대조구에 비해 발근수와 뿌리길이에서 약 2.1배의 뿌리 신장 효과를 보였고, 상추발근 생검법에서는 대조구에 비해 뿌리길이와 무게에서 약 1.8배의 뿌리신장효과를 보였다.

This study investigated the synergistic effect of single inoculation and co-inoculation of phosphate-solubilizing bacteria (PSB) Burkholderia metallica JH-7 and Burkholderia contaminans JH-15. Phosphate-solubilizing abilities of these strains were assessed by measuring phosphorus content in culture media that were singly inoculated or co-inoculated with these strains for 7 days. B. metallica JH-7 was found to release the highest content of soluble phosphorus (140.80 μg mL-1) into the medium, followed by single inoculation of B. contaminans JH-15 (135.95 μg mL-1) and co-inoculation of two strains (134.84 μg mL-1). The highest pH reduction, organic acid production, and glucose consumption were observed in the medium inoculated with B. metallica JH-7 alone compared with that in the medium co-inoculated with both the strains. Results of a plant growth promotion bioassay showed 17.4% and 7.48% higher leaf and root growth, respectively, in romaine lettuce inoculated with B. metallica JH-7 alone than in romaine lettuce inoculated with a control strain. However, no significant difference was observed between single inoculation and co-inoculation of these strains with respect to phosphorus release and plant growth. Although the results of the present study did not show the synergistic effect of phosphate solubilization by the PSB strains examined, these results indicate that treatment with PSB exerts a beneficial effect on crop growth.

양송이 수확후 배지로부터 lipase 생산균을 분리하여 16S rDNA 유전자 분석을 통해 동정한 결과, Burkholderia cepacia ATCC와 99.8% 상동성을 나타냈다. 분리균 B. cepacia 배양여액 중에 함유된 효소단백질을 70% 황산암모늄으로 침전시켜 crude lipase를 회수하였다. 고정화 효소를 제조하기 위하여 crude lipase(CL)과 Novozyme lipase(NL)을 cross-linking 법에 의해 Silane화된 Silicagel에 고정화 시킨 결과, immobilized CL(ICL)은 61%, immobilized NL(INL)은 72%의 잔존활성을 유지하였다. 중성지방 Canola oil을 알칼리(NaOH) 촉매와 효소(CL 및 ICL) 촉매를 이용하여 지방산(fatty acid)으로 분해한 후, methanolysis에 의한 에스터전이반응(trans-esterification)을 통해 지방산으로부터 전환된 바이오디젤(fatty acid methyl ester, FAME)의 종류와 수율을 비교 하였다. 생성된 총 FAME 함량은 NaOH 781mg L-1, free lipase 681mg L-1, 고정화 lipase 598mg L-1순으로 높았으며, 지방산 조성별 FAME 함량은 linoleic acid(C18:1)가 약 50%로 가장 높았으며, stearic acid(C18:0)가 22%정도의 높은 수준이었다. 또한 반응시간이 증가함에 따라 CL과 ICL 모두 불포화지방산 FAME의 조성비는 감소하고, 상대적으로 포화지방산 FAME의 조성비는 증가하는 경향을 보여 lipase 효소가 transesterification 활성과 interesterification 활성을 동시에 가지는 것으로 여겨진다. 고정화효소의 잔여활성은 반복회수가 증가함에 따라 서서히 감소하여 4회 반복 후, 초기 활성도에 비해 ICL은 34% 와 INL은 21%까지 감소하였다.

A galactose fermentation bacterium producing lactose from red seaweed, which was known well to com-promise the galactose as main reducing sugar, was isolated from button mushroom bed in Buyeo-Gun, Chungchugnam-do province. The lactic acid bacteria MONGB-2 was identified as Lactobacillus paracasei subsp. tolerans by analysisof 16S rRNA gene sequence. When the production of lactic acid and acetic acid by L. paracasei MONGB-2 was inves-tigated by HPLC analysis with various carbohydrates, the strain MONGB-2 efficiently convert the glucose and galactoseto lactic acid with the yield of 18.86 g/L and 18.23 g/L, respectively and the ratio of lactic acid to total organic acidswas 1.0 and 0.91g/g for both substrates. However, in the case of acetic acid fermentation, other carbohydrates besidesgalactose and red seaweed hydrolysate could not be totally utilized as carbon sources for acetic acid production by thestrain. The lactic acid production from glucose and galactose in the fermentation time courses was gradually enhancedupto 60 h fermentation and the maximal concentration reached to be 16-18 g/L from both substrates after 48 h of fer-mentation. The initial concentration of glucose and galactose were completely consumed within 36 h of fermentation, ofwhich the growth of cell also was maximum level. In addition, the bioconversion of lactic acid from the red seaweedhydrolysate by L. paracasei MONGB-2 appeared to be about 20% levels of the initial substrates concentration and thisresults were entirely lower than those of galactose and glucose showed about 60% of conversion. The apparent resultsshowed that L. paracasei MONGB-2 could produce the lactic acid with glucose as well as galactose by the homof-ermentation through EMP pathway

Y-Aminobutyric acid(GABA) is a four carbon non-protein amino acid that has several well-known phys-iological functions, such as a postsynaptic inhibitory neurotransmitter in the brain and induction of hypotensive and tran-quilizer effects. A lactic acid bacterium was isolated from button mushroom bed, which is showing high GABAproductivity by TLC or HPLC analysis. The strain was identified as Lactobacillus hilgardii by analysis of 16S rDNAgene sequence. When the maximum production of GABA by L. hilgardii was investigated with various concentration ofmonosodium glutamate, the yield of GABA reached to be 53.65mM at 1% mono sodium glutamate (MSG) in flask cul-tivation. A Glutamate decarboxylase (GAD) enzyme, which was known to convert MSG to GABA, was purified froma cell-free extract of L. hilgardii and the molecular weights of purified GAD was estimated to 60,000 by SDS-PAGE.The optimum pH and temperature of GAD were at pH4.6 and at 37°C, respectively. The GAD activity was increasedby the addition of sulfate ions such as ammonium sulfate, sodium sulfate and magnesium sulfate, indicating that theincrease of hydrophobic interaction causes the increase of GAD activity.